Species Diagnostics Protocols: Pcr And Other Nucleic Acid Methods

CHAPTER 1 Isolation and Purification of Plant Nucleic Acids Genomic and Chloroplast Mien T. G. van de Ven, Patrick and Rex M. Brennan DNA G. Lanham, 1. Introduction The use of molecular protocols has expanded into virtually all branches of plant science in recent years, and crucial to theseprotocols is the effective isolation of plant nucleic acids in a purified state. Isolation of DNA from plant tissue must be simple, rapid, inexpensive, reproducible, and efficient, particularly when many samples are required, e.g., in population studies. The increasing use of polymerase chain reaction (PCR)based technology in plant molecular biology, especially in transgenic analysis, genetic mapping for genome analysis, and genetic fingerprinting, has placed further emphasis on the need for the efficient isolation of pure DNA, often from small amounts of plant tissue. Isolation of highly purified plant DNA is often complex, particularly from plant tissues high in polyphenolic compounds that can react with cellular enzymes during the extraction procedure to render the DNA unsuitable for further analysis. Contamination with polysaccharides, mainly pectins, is also a frequent problem; the following protocols are designed to limit their effects. Protocols for both genomic and chloroplast DNA (cpDNA) are given. CpDNA is now widely used, particularly in association with restriction From Methods Nuclerc In Molecular Acrd Methods B/ology, Edlted Vol 50. Species Dlagnostm Protocols PCR and Other by. J P Clapp Humana Press Inc , Totowa, NJ 1 van de V